LOCUS       sequence-249311-        6422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PPI (BiFC/TriFC) assay vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6422)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6422)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pRDAX-466    N-Tag UnaG[2] L20 FKBP mCherry
FEATURES             Location/Qualifiers
     source          1..6422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        533..736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     687..707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     733..757
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          872..1004
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1019..1028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1253..1576
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(1590..2164,2165..2167,2168..2176)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1757..1774)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1984..2003
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2177..2887
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2322..2340)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2590..2609
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2933..3065
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3074..3662
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3127..3147)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3545..3556)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3689..3706)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3695..3806
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3902..3922)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3902..3920)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3941..3957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3941..3957)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3954..3976)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4059..4078)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4124..4179
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4178..4197)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4558..5146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4638..4657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5317..6108,6109..6177))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5940..5959
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6178..6282)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6350..6368
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcctgcctcc aggcctcaat attggccatt agccatatta ttcattggtt atatagcata
       61 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta
      121 tattggctca tgtccaatat gaccgccatg ttggcattga ttattgacta gttattaata
      181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      361 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
      421 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
      481 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      541 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      601 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      661 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
      721 ctatataagc agagctcgtt tagtgaaccg tcagatcact agaagcttta ttgcggtagt
      781 ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct cgaacttaag
      841 ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
      901 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
      961 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccaggc
     1021 caccatgggg gtgaagtctg tggtgaatct ggtaggcgag aaactcgtgt acgtccagaa
     1081 gtgggatggg aaggaaacca cgtatgtgcg agagatcaag gacgggaagt tggttgtcac
     1141 cctgaccatg ggcgatgtgg ttgctgtgcg ctcatatcgg agagccactg aagggagtcc
     1201 tggcggtgga tccggaggct ctgggagctc tgccagcggt ggcagcacct ccggagtgca
     1261 ggtggaaacc atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgcgt
     1321 ggtgcactac accgggatgc ttgaagatgg aaagaaattt gattcctccc gggacagaaa
     1381 caagcccttt aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt
     1441 tgcccagatg agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg
     1501 tgccactggg cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct
     1561 tctaaaactg gaataagaat tcagatccgc ccctctccct cccccccccc taacgttact
     1621 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     1681 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     1741 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     1801 gcagttcctc tggtagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     1861 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     1921 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     1981 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     2041 tgtatgggat ctgatctggg gcctcggtac acatgcttta catgtgttta gtcgaggtta
     2101 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     2161 taatatggcc acaaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     2221 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     2281 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     2341 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     2401 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     2461 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     2521 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     2581 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     2641 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     2701 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     2761 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     2821 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     2881 caagtaaacc ggtgctcgag gtaccgatga atagctaagg tcgaggccgc aggtaagtat
     2941 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
     3001 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
     3061 cacaggtgtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3121 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3181 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3241 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3301 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3361 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3421 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     3481 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3541 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3601 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3661 gcctggaatt cgagctcggt acgatcagcc tcgactgtgc cttctagttg ccagccatct
     3721 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3781 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3841 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagccc agcttttgtt
     3901 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3961 gaaattgtta tccgctaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     4021 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     4081 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     4141 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     4201 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     4261 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     4321 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     4381 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     4441 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     4501 agatcggccg gccgccggcg tcgacgcgcg cggccgcggc gcgccgttgc tggcgttttt
     4561 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4621 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4681 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4741 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4801 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4861 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4921 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4981 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5041 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5101 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5161 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5221 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5281 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5341 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5401 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5461 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5521 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5581 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5641 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5701 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5761 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5821 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5881 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5941 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6001 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6061 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6121 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6181 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6241 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6301 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6361 cacgaggccc tttcgtctac gcgcgcgccg gcggcgcgcc gtcgacggcc ggccgcggcc
     6421 gc
//
